DOTAP chloride
(Synonyms: (2,3-二油氧基丙基)三甲基氯化铵,1,2-Dioleoyl-3-trimethylammoniumpropane (chloride)) 目录号 : GC41811
DOTAP chloride 是一种脂质体制剂，已用于将质粒 DNA 转染到培养的真核细胞中 [1]。
Cas No.:132172-61-3
Sample solution is provided at 25 µL, 10mM.
;)
,-1-7$$$37316672.jpg');)
;)
;)
$$$36806353.jpg');)
;33(7)%EF%BC%9A546-561$$$37156877.jpg');)
;)
;)
;)
%201124-1138.$$$35908550.jpg');)
%20766-780.$$$37100057.jpg');)
%20418-432.$$$36893736.jpg');)
%EF%BC%9A-240-255.$$$35108512.jpg');)
533-545$$$36543525.jpg');)
%201-13.$$$36600053.jpg');)
;)
DOTAP chloride is a liposome formulation that have been used to transfect plasmid DNA into eukaryotic cells in culture [1]. The synthetic cationic amphiphile DOTAP chloride has been shown to mediate uptake of plasmid also in the presence of a culture medium containing serum. It is metabolized in the cells and has proven to be less cytotoxic than other amphiphiles [2]. DOTAP chloride has also been used successfully for internalization of an 18-mer DNA oligonucleotide, and it was shown in these experiments to protect the oligonucleotide from degradation by nucleases in serum and cells [3].
References:
[1]. Haukenes G, Szilvay A M, Brokstad K A, et al. Labeling of RNA transcripts of eukaryotic cells in culture with BrUTP using a liposome transfection reagent (DOTAP?)[J]. Biotechniques, 1997, 22(2): 308-312.
[2]. Leventis R, Silvius J R. Interactions of mammalian cells with lipid dispersions containing novel metabolizable cationic amphiphiles[J]. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1990, 1023(1): 124-132.
[3]. Capaccioli S, Dipasquale G, Mini E, et al. Cationic lipids improve antisense oligonucleotide uptake and prevent degradation in cultured cells and in human serum[J]. Biochemical and biophysical research communications, 1993, 197(2): 818-825.
DOTAP chloride 是一种脂质体制剂，已用于将质粒 DNA 转染到培养的真核细胞中 [1]。合成的阳离子两亲物 DOTAP chloride 已被证明在含有血清的培养基存在下也能介导质粒的摄取。它在细胞内代谢，已证明其细胞毒性低于其他两亲物 [2]。 DOTAP 氯化物也已成功用于 18 聚体 DNA 寡核苷酸的内化，并且在这些实验中表明它可以保护寡核苷酸免于被血清和细胞中的核酸酶降解[3]。
| Cell experiment [1]: | |
|
Cell lines |
mammalian CV-1 and 3T3 cells |
|
Preparation Method |
Lipid dispersions containing cationic lipid analogues were prepared by first bath-sonicating dried lipid samples in distilled water to clarity (2-5 min), then adding an equal concentration of 308 mM NaCl, 40 mM Hepes (pH 7.4), and sonicating further for 2 min. Lipid/DNA mixtures for transfections were prepared by incubating dotap chloride with DNA for 5 min, typically at a total lipid concentration of 0.5-2 mg/ml, before addition to cell monolayers. Reverse-phase evaporation vesicles were prepared as described previously [2] and filtered through 0.1/xm pore-size Nucleopore filters. Lipid mixing between cationic lipid dispersions and large unilamellar PC/PS vesicles was assayed by a resonance energy transfer-based procedure as described previously [3], incubating cationic lipid dispersions (2 μM), colabeled with 1 mol% (12-CPS)-18 PC and 0.35 mol% (12-DABS)-18 PC, with a 9-fold excess (18 μM) of unlabeled PC/PS vesicles. For transfection of cells using cationic lipid dispersions, cell monolayers were washed twice with HEPESbuffered saline, then incubated with lipid/DNA mixtures in the latter medium (total volume 3 ml per100-mm culture dish) at 37 °C, for 90 min except whereotherwise indicated. Following this incubation period,the incubation medium was either replaced or supplemented with 12 ml of D-MEM containing 5% serum,and the cells were cultured for a further 36 h beforeharvesting. Chloramphenicol acetyltransferase activityin plasmid pSV2cat-transfected cells was assayed asdescribed previously [4], using chloramphenicol andacetyl-CoA concentrations of 3.5 μM and 2.5 mM,respectively. |
|
Reaction Conditions |
0.5-2 mg/ml for 90 min |
|
Applications |
For transfection of the cells with a fixed amount of plasmid (usually 2 or 5 µg) |
|
References: [1]: Leventis R, Silvius J R. Interactions of mammalian cells with lipid dispersions containing novel metabolizable cationic amphiphiles[J]. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1990, 1023(1): 124-132. [2]: Wilschut J, Duzgunes N, Fraley R, et al. Studies on the mechanism of membrane fusion: kinetics of calcium ion induced fusion of phosphatidylserine vesicles followed by a new assay for mixing of aqueous vesicle contents[J]. Biochemistry, 1980, 19(26): 6011-6021. [3]: Silvius J R, Leventis R, Brown P M, et al. Novel fluorescent phospholipids for assays of lipid mixing between membranes[J]. Biochemistry, 1987, 26(14): 4279-4287. [4]: Gorman C M, Moffat L F, Howard B H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells[J]. Molecular and cellular biology, 1982, 2(9): 1044-1051. |
|
| Cas No. | 132172-61-3 | SDF | |
| 别名 | (2,3-二油氧基丙基)三甲基氯化铵,1,2-Dioleoyl-3-trimethylammoniumpropane (chloride) | ||
| Canonical SMILES | CCCCCCCC/C=C\CCCCCCCC(OC(C[N+](C)(C)C)COC(CCCCCCC/C=C\CCCCCCCC)=O)=O.[Cl-] | ||
| 分子式 | C42H80NO4•Cl | 分子量 | 698.6 |
| 溶解度 | ≤33mg/ml in ethanol;0.5mg/ml in DMSO;5mg/ml in dimethyl formamide | 储存条件 | Store at -20°C, protect from light, stored under nitrogen |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
||
| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
 |
1 mg | 5 mg | 10 mg |
| 1 mM | 1.4314 mL | 7.1572 mL | 14.3143 mL |
| 5 mM | 0.2863 mL | 1.4314 mL | 2.8629 mL |
| 10 mM | 0.1431 mL | 0.7157 mL | 1.4314 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Quality Control & SDS
- View current batch:
- Purity: >99.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet